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Dynamic single-cell phenotyping of immune cells using the microfluidic platform DropMap.

Yacine Bounab1,2, Klaus Eyer3,4, Sophie Dixneuf5

  • 1BIOASTER Technology Research Institute, Lyon, France.

Nature Protocols
|August 14, 2020
PubMed
Summary
This summary is machine-generated.

A new microfluidic platform, DropMap, enables dynamic, quantitative analysis of single immune cells and their secreted proteins like cytokines and antibodies, overcoming limitations of current methods.

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Area of Science:

  • Immunology
  • Microfluidics
  • Cell Biology

Background:

  • Current methods for immune response characterization lack quantitative, dynamic analysis of single cells and secreted proteins.
  • Existing techniques like ELISPOT, flow cytometry, CyTOF, and single-cell sequencing provide only end-point measurements.
  • There is a need for systems that can simultaneously measure secreted protein kinetics and cellular phenotypes.

Purpose of the Study:

  • To introduce and validate DropMap, a microfluidic platform for high-throughput, quantitative, and dynamic analysis of single immune cells.
  • To demonstrate the platform's capability in measuring secretion kinetics of cytokines and antibodies alongside cellular characteristics.
  • To showcase DropMap's application in profiling immune responses in disease contexts.

Main Methods:

  • Development of a microfluidic platform (DropMap) for compartmentalizing single cells in 50-pL droplets.
  • Integration of fluorescence microscopy with a paramagnetic nanoparticle-based immunoassay for quantitative protein detection.
  • Simultaneous measurement of secreted protein kinetics, endocytosis activity, viability, and cell-surface markers.

Main Results:

  • DropMap enables simultaneous measurement of secretion kinetics and multiple cellular phenotypes from tens of thousands of single immune cells.
  • The protocol is robust, with a total assay time of 8-10 hours.
  • Demonstrated applications include profiling tumor necrosis factor-α (TNF-α) secretion in septic shock patients, analyzing T cell cytokine secretion rates, and measuring B cell antibody affinity.

Conclusions:

  • DropMap offers a significant advancement for characterizing immune responses by enabling quantitative, dynamic, single-cell analysis of secreted proteins.
  • The platform overcomes limitations of existing end-point measurement techniques.
  • DropMap has broad applicability for studying immune cell function in health and disease.